Diagnosis of degenerative collagen-related skin diseases is a complex process that requires histopathological evaluation of various pathological alterations, which in addition are not unambiguous in themselves. Fluorescence spectroscopy has proven to be valuable tool for tissue differentiation, whereas the biomedical application of tissue polarimetry is establishing as a valuable diagnostic modality. In this work we present the evaluation of experimental results of histology tissue slides from three collagen-related skin diseases: psoriasis, lupus and scleroderma, through fluorescence spectroscopy and Muller polarimetry.
We investigated pigmented skin tumour lesions in vivo and ex vivo, including benign and dysplastic nevi, as well as malignant lesions, such as pigmented basal cell carcinoma (BCC) and malignant melanoma (MM) lesions, to obtain a complex view about the feasibility of different excitation sources solely and/or in combination to induce fluorescence signal useful for diagnosis of various low-fluorescent cutaneous neoplasia. A specialized multispectral analysis of the data obtained was applied by using excitation in broad spectral range, covering ultraviolet, visible and near-infrared spectral range, that contribute considerably to: (1) fundamental determination of tumour tissues’ spectral properties, and (2) to increase the accuracy in determining the type of cutaneous pathology. The chromophores, related to the formation of ultraviolet and visible (UV-VIS) fluorescence in human normal skin and its pigmented lesions are mainly amino acids – tryptophan, tyrosine; structural proteins and their cross-links – collagen, elastin, keratin; co-enzymes - NADH, flavins; vitamins and lipids. In the near-infrared (NIR) spectral region, skin fluorescence emission properties are related to the presence of melanin pigment, lipids and endogenous porphyrins, if any, as the highest impact on the resultant emission spectrum is due to the melanin compound.
We investigated melanin-pigmented skin samples ex vivo, of benign and dysplastic nevi, as well as malignant melanoma, obtained after surgical excision, containing so called safety areas, where a normal skin could be observed, to obtain a complex and complete view about the feasibility of different excitation sources solely and/or in combination to induce fluorescence signal useful for diagnosis of pigmented cutaneous neoplasia. Using the specialized multispectral analysis of the data obtained by fluorescence using excitation in broad spectral range, covering ultraviolet, visible and near-infrared spectral range, contribute considerably to the both the fundamental determination of tumour tissues’ basic spectral parameters, and to increasing the accuracy and specificity in determining the type of pathology when spectral techniques are used that are applicable to the clinical practice. Excitation wavelengths applied namely were 365, 385, 405, 630, 785 nm. As excitation sources were used set of laser and light emitting diodes with output power in the frames of 20-50mW. Microspectrometers USB4000 and QE65000 (Ocean Optics Inc, USA) in the range of 350-1100 nm were used as detectors of the fluorescence spectra obtained using UV-VIS-NIR excitation. These are preliminary investigations based on ex vivo samples, which would be followed by in vivo investigations and comparative studies to obtain the diagnostic accuracy, using optical spectral techniques, which would surpass the existing clinical techniques for early detection and evaluation of skin cancer. The achieved results demonstrate autofluorescence spectra efficiency in highlighting biochemical changes during tumor growth in a broad spectral range.
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